Nucleotide molecule SR2B2 and application of the same in producing antidiabetic medicament
An anti-diabetic and nucleotide technology, applied in the field of biomedicine, can solve the problems of gene drugs that have not been reported against diabetes, and achieve the effect of promoting insulin secretion
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Embodiment 1
[0031] Example 1 Effect of siRNA on insulin secretion after interfering with endogenous BRSK2 expression
[0032] (1) Dilute NIT cells at 1×10 5 Cells / well were seeded on a 24-well plate, and the cells were placed in CO 2 Cultivate in an incubator at 37° C. for 18-24 hours to make the cell abundance reach 70-80%.
[0033] (2) Invitrogen lipofectamine 2000 eukaryotic transfection kit was used for transfection. 5 μl of Nonsilence (the negative control of siRNA interference), siRNA (GCUGCACGACGUUUAUGAA dTdT and dTdTUUCAUAAACGUCGUGCAGC)-51 μl, 3 μl and 5 μl (the concentration of each μl is 20 μM) were transfected respectively. Four wells were replicated for each transfection.
[0034](3) 16 hours after transfection, replace with complete medium (DMEM high glucose medium + 15% fetal calf serum) and continue to culture for 24 hours.
[0035] (4) The complete medium (sugar concentration: 25 mM) was replaced again, and the culture supernatant was collected after culturing for 20 m...
Embodiment 2
[0041] Example 2 Effect of BRSK2 overexpression on insulin secretion
[0042] (1) Dilute NIT cells at 1×10 5 Cells / well were seeded on a 24-well plate, and the cells were placed in CO 2 Cultivate in an incubator at 37° C. for 18-24 hours to make the cell abundance reach 70-80%.
[0043] (2) Invitrogen lipofectamine 2000 eukaryotic transfection kit was used for transfection. Two plasmids, Pcmv-Myc empty vector and Pcmv-Myc-BRSK2, were transfected respectively, and each plasmid was transfected into 4 wells at a concentration of 200ng / well.
[0044] (3) 16 hours after transfection, replace with complete medium (DMEM high glucose medium + 15% fetal calf serum) and continue to culture for 24 hours.
[0045] (4) The complete medium (sugar concentration: 25 mM) was replaced again, and the culture supernatant was collected after culturing for 20 minutes.
[0046] (5) The concentration of insulin in the culture supernatant was measured by radioimmunoassay. The detection kit was pro...
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